Separation of Large DNA Fragments by Capillary Electrophoresis under Pulsed-Field Conditions

Abstract

Capillary zone electrophoresis (CZE), using an entangled polyacrylamide solution, was applied to large DNA samples under pulsed-field conditions. Highly efficient separations were achieved under biased sinusoidal field and field-inversion pulsing regimes. The separations that were obtained with 8.3-48.5 kb lambda DNA standards and 48.5 kb-1 Mb lambda DNA concatamers (modified with ethidium bromide) clearly demonstrate a dramatic improvement in the separation time (roughly, 10-50 times) over the conventionally used slab-gel techniques. Moreover, the CZE method appears much more sensitive and amenable to component quantification and method automation.